Nongenomic action and mechanism of 17β-estradiol in cytosolic calcium concentration in delayed implantation mouse endometrial stromal cells

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WANG Qiang(Department of Physiology, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China)
YUE Li-Min(Department of Physiology, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China)
ZHANG Jin-Hu(Department of Physiology, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China)
TIAN Ji-Mei(Department of Physiology, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China)
HE Ya-Ping(Department of Physiology, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China)
Journal Title:
Volume 60, Issue 02, 2008
Key Word:
estrogen; phospholipase C; calcium; stromal cells

Abstract: To investigate the existence of nongenomic action of 17β-estradiol (E2) in the delayed implantation mouse endometrialstromal ceils, the changes in intraceilular calcium concentration ([Ca2+]I) and the upstream of calcium signal in vitro were detected. Theexperiment was composed of two parts. Firstly, the change in [Ca2+]I in endometrial stromal cells induced by E2 under differentconditions was detected. The mice were divided into 6 groups as follows: 0.1% dimethylsulfoxide (DMSO) control group, 1×10-8 mol/Lbovine serum albumin (BSA) control group, 1×10-8 mol/L E2 group, 1×10-8 mol/L E2 conjugated with BSA (E2-BSA) group, 1×10-8 mol/LE2 + calcium-free medium group, 1×10-8 mol/L E2 + 5μg/mL tamoxifen group, with 4 mice in each group. The endometrial tissue wasobtained from delayed implantation mice at pregnant day 7, and digested by incubation of tissue minces in Hank's balanced salts(HBSS, pH 7.2), which contained glucose (1 g/L), and eollagenase I (0.125%), for 1 h at 37 ℃. The stromal cells were preloaded with2.5 mmol/L Fluo-3/AM, a fluorescent probe of calcium, for 30 rain. A confocal laser scanning microscope, which fixed the wave lengthof excitation and emission at 488 nm and 526 nm, respectively, was used to detect the change in [Ca2+]I Secondly, the mechanism of E2effects in endometrial stromal cells was investigated. Immunofluorescent method was used to detect the change in phosphorylation ofphospholipase C (PLC) before and after the stromal cells were treated with E2 for 5 rain, 15 rain, and 30 rain. Seven delayedimplantation mice were used. The results were as follows. [Ca2+]I increased immediately and reached the maximum at 15 rain after the stromal cells were treated with 1×10-8 mol/L E2 and returned to the normal level at 30 min. In E2-BSA group and E2 + calcium-free medium group the same results were obtained as that in E2 group, but there was no increase of [Ca2+]I in DMSO and BSA groups. Tamoxifen, a traditional antagonist of estrogen receptor, did not inhibit the increase in [Ca2+]I induced by E2. Immunofluorescent results showed that the change in phosphorylated-PLC level had the same trend as [Ca2+]I after the cells were treated with E2. Compared with that in the control group, the immunofluorescent intensity increased at the beginning and achieved the maximum at 15 min (P<0.001), then declined to the normal level at 30 min. These results suggest that the rapid response of [Ca2+]I induced by E2 in the endometrial stromal cells in delayed implantation mice is possibly carded out through a nongenomic pathway, and the transmembrane signal transduction is related to the phosphorylation of PLC in this process.

  • [1]Dey SK,Lim H,Das SK,Resse J,Paria BC,Daikoku T,Wang HB.Molecular cues to implantation.Endocr Rev 2004; 25(3):341-373.
  • [2]Paria BC,Lim H,Das SK,Resse J,Dey SK.Molecular signaling in uterine receptivity for implantation.Cell Dev Biol 2000; 11:67-76.
  • [3]Paria BC,Reese J,Das SK,Dey SK.Deciphering the crosstalk of implantation:advances and challenges.Science 2002; 296:2185-2188.
  • [4]Conneely OM.Perspective:female steroid hormone action.Endocrinology 2001; 142(6):2194-2199.
  • [5]Hall JM,Couse JF,Korach KS.The multifaceted mechanisms of estradiol and estrogen receptor signaling.J Biol Chem 2001;276 (40):36869-36872.
  • [6]Revelli A,Massobrio M,Tesarik J.Nongenomic actions of steroid hormones in reproductive tissues.Endocr Rev 1998;19(1):3-17.
  • [7]Luconi M,Francavilla F,Porazzi I,Macerola B,Forti G,Baldi E.Human spermatozoa as a model for studying membrane receptors mediating rapid nongenomic effects of progesterone and estrogens.Steroids 2004; 69:553-559.
  • [8]Tesarik J,Mendoza C.Nongenomic effects of 17β-estradiol on maturing human oocytes:relationship to oocyte developmental potential.J Clin Endocrinol Metab 1995; 80 (4):1438-1443.
  • [9]Younglai EV,WU YJ,Kwan TK,Kwan CY.Non-genomic action of estradiol and progesterone on cytosolic calcium concentrations in primary cultures of human granulosa-lutein cells.Hum Reprod 2005; 20 (9):2383-2390.
  • [10]Wang TH (王庭槐),Fu XD,Yang D,Tan Z,Pan JY.Membrane estrogen receptor mediates the rapid nongenomic activation of endothelial nitric oxide synthase by estrogen.Acta Physiol Sin (生理学报) 2003; 55(2):213-218 (Chinese,English abstract).
  • [11]Paria BC,Huet-Hudson YM,Dey SK.Blastocyst's state of activity determines the "window" of implantation in the receptive mouse uterus.Proc Natl Acad Sci USA 1993; 90:10159-10162.
  • [12]Yu LL (喻琳麟),Hu Y,Zhang JH,He YP,Yue LM.Rapid effects of estrogen on intracellular calcium of mouse dormant blastocysts.Chin J Cell Biol (细胞生物学杂志) 2007; 29(4):569-572 (Chinese,English abstract).
  • [13]Cooke PS,Buchanan DL,Lubahn DB,Cunha GR.Mechanism of estrogen action:lessons from the estrogen receptor-α knockout mouse.Biol Reprod 1998; 59:470-475.
  • [14]Cooke PS,Buchanan DL,Young P,Setiawan T,Brody J,Korach KS,Taylor J,Lubahn DB,Cunha GR.Stromal estrogen recepors mediate mitogenic effects of estradiol on uterine epithelium.Proc Natl Acad Sci USA 1997; 94:6535-6540.
  • [15]Ma WG,Song H,Das SK,Paria BC,Dey SK.Estrogen is a critical determinant that specifies the duration of the window of uterine receptivity for implantation.Proc Natl Acad Sci USA 2003; 100 (5):2963-2968.
  • [16]Pietras RJ,Szego CM.Endometrial cell calcium and oestrogen action.Nature 1975; 253:357-359.
  • [17]Kayisli OG,Kayisli UA,Luleci G,Arici A.In vivo and in vitro regulation of akt activation in human endometrial cells is estrogen dependent~ Biol Reprod 2004; 71:714-721.
  • [18]Berridge MJ,Lipp P,Bootman MD.The versatility and universality of calcium signalling.Nat Mol Cell Bio12000; 1:11-21.
  • [19]Fu JD (傅继东),Yang HT.Developmental regulation of intracellular calcium homeostasis in early cardiac myocytes.Acta Physiol Sin (生理学报) 2006; 58(2):95-103 (Chinese,English abstract).
  • [20]Berridge M J,Galione A.Cytosolic calcium oscillators.J FASEB 1988; 2:3074-3082.
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